GP350 AS A NOVEL VACCINE PROTEIN CARRIER

GP350 作为新型疫苗蛋白载体

• 批准号: 7958394
• 负责人: CLIFFORD M SNAPPER
• 金额: $ 6.49万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7958394
• 关键词: Adjuvant; Adolescent; Affinity; Antibodies; B-Lymphocytes; Binding; Biological Assay; CD4 Positive T Lymphocytes; Carrier Proteins; Clinical Trials; Complement 3d Receptors; Complement Receptor; Computer Retrieval of Information on Scientific Projects Database; Data Analyses; Dose; Engineering; Enzyme-Linked Immunosorbent Assay; Epstein-Barr Virus Infections; Follicular Dendritic Cells; Funding; Goals; Grant; Hodgkin Disease; Human; Human Herpesvirus 4; Immunization; In Vitro; Incidence; Infant; Infection; Infectious Mononucleosis; Institution; Macaca mulatta; Mediating; Mus; N-terminal; Nasopharynx Carcinoma; New England; Non-Hodgkin' s Lymphoma; Plasma; Polysaccharides; Population; Primates; Proteins; Recombinants; Recruitment Activity; Relative (related person); Research; Research Design; Research Personnel; Resources; Sampling; Serotyping; Source; Syndrome; T-Lymphocyte Epitopes; United States National Institutes of Health; Vaccines; Vertebral column; Viral Proteins; aluminum sulfate; anti-IgG; base; cell transformation; immunogenic; meetings; neutralizing antibody; nonhuman primate; novel vaccines; preclinical study; response; young adult

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There currently exists a need for a vaccine that is protective against Epstein-Barr virus (EBV) infection. Such a vaccine would have a potential global impact on the incidence of infectious mononucleosis, Hodgkin's and non-Hodgkin's lymphoma, nasopharyngeal carcinoma, and lymphoproliferative syndrome. The EBV protein, gp350 has the potential to meet this need. Thus, 1) gp350 binds to the complement receptor type 2 (CD21) on B cells and follicular dendritic cells (FDCs) and thus may act as a potent adjuvant, 2) gp350 is the main target for antibody-mediated EBV neutralization, 3) multimerization of gp350 on a PS backbone will itself enhance anti-gp350 responses, and 4) gp350 is an immunogenic foreign protein that will recruit CD4+ T cell help for induction of anti-gp350 antibody. In the developing world infection with EBV, occurs in the infant population. Since, in contrast, peak EBV infection in the developed world occurs in adolescents, gp350 would also be of use as an anti-EBV vaccine in this latter population. In preliminary studies we have: 1) expressed and purified a recombinant glycosylated N-terminal 72kDa fragment of the gp350 molecule, 2) conjugated multiple copies of gp350 to pneumococcal capsular polysaccharide, serotype 14 (PPS14) [PPS14-gp350], 3) demonstrated the ability of PPS14-gp350 to specifically bind to CD21 expressed on rhesus and human, but not murine, B cells, and 4) induced boosted plasma IgG anti-PPS14 and IgG anti-gp350 antibodies in young adult rhesus monkeys following i.m. immunization with as little as 0.05 mg of PPS14-gp350 adsorbed on alum. However, careful analysis of the data, and EBV neutralizing ability of the plasma samples led us to hypothesize that a new, genetically engineered multimeric gp350 containing strong T cell epitopes would be a superior vaccine. The goal of this proposal is to extend the above studies to determine, using rhesus monkeys, the potential ability of a new genetically engineered multimeric gp350 to serve as a highly potent EBV vaccine . These pre-clinical studies will form the basis for progressing directly to human clinical trials. In this proposal we will utilize young adult rhesus monkeys to: Directly compare the ability of two distinct doses of alum-adsorbed monomeric or genetically engineered multimeric gp350 to a. elicit primary, secondary, and tertiary plasma IgG anti-gp350 titers and induce affinity maturation as determined by ELISA. b. induce EBV-neutralizing antibody as determined by both an in vitro B cell transformation and ELISA neutralization assay. c. prime specific CD4+ T cells as determined by an in vitro re-stimulation assay. Collectively, these studies are designed to establish a proof-of-principle in non-human primates, for using gp350 clinically as a protective, antibody-based vaccine for EBV relative to monomeric gp350.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 目前，需要一种疫苗，该疫苗可抵抗爱泼斯坦 - 巴尔病毒（EBV）感染。这样的疫苗将对传染性单核细胞增多症，霍奇金和非霍奇金淋巴瘤，鼻咽癌和淋巴结增生性综合征的发生率产生潜在的全球影响。 EBV蛋白GP350有可能满足这种需求。因此，1）GP350与B细胞和卵泡树突状细胞（FDC）上的补体2型（CD21）结合，因此可以充当有效的佐剂，2）GP350是抗体介导的EBV中和的主要靶标，是抗体介导的EBV中和的主要靶标，3）ps Backbone上的GP350响应于pp350的GP350，抗gp350响应于pp350 gp350 gp3 50，4这将募集CD4+ T细胞帮助诱导抗GP350抗体。在发展中世界的EBV感染中，婴儿人群发生。相比之下，由于发达国家发生EBV峰值感染发生在青少年中，因此在后一种人群中，GP350也将用作抗EBV疫苗。 在初步研究中，我们有：1）表达并纯化了GP350分子的重组糖基化的N末端72KDA片段，2）共轭GP350的多个副本，将GP350的多个拷贝与肺炎球菌囊囊多糖糖糖糖糖含量，血清型14（PPS14）[PPS14-gp3 50]，PPP-gp3 50 pp，3），3） CD21在恒河猴和人类，但不表示鼠，B细胞和4）诱导的年轻成人恒河猴的促进血浆IgG抗PPPS14和IgG抗GP350抗体之后I.M.免疫，几乎只有0.05 mg的PPS14-GP350吸附在明矾上。但是，对数据的仔细分析以及血浆样品的EBV中和能力导致我们假设一种新的，基因工程的多聚体GP350含有强T细胞表位是一种优越的疫苗。 该提案的目的是扩展上述研究，以确定新的基因工程多聚体GP350作为一种高度有效的EBV疫苗的潜在能力。这些临床前研究将构成直接进入人类临床试验的基础。在此提案中，我们将利用年轻的成年恒河猴来： 直接将两种不同剂量的明矾吸附的单体或基因工程多聚体GP350与 一个。 引起原发性，次级和第三级IgG抗GP350滴度，并诱导ELISA确定的亲和力成熟。 b。 通过体外B细胞转化和ELISA中和测定法确定的EBV中和抗体。 c。 由体外重新刺激测定法确定的主要特异性CD4+ T细胞。 总的来说，这些研究旨在在非人类灵长类动物中建立原则证明，用于在临床上使用GP350作为一种基于保护性的，基于抗体的疫苗，用于EBV相对于单体GP350。